New ABA-Hypersensitive Arabidopsis Mutants Are Affected in Loci Mediating Responses to Water Deficit and Dickeya dadantii Infection

On water deficit, abscisic acid (ABA) induces stomata closure to reduce water loss by transpiration. To identify Arabidopsis thaliana mutants which transpire less on drought, infrared thermal imaging of leaf temperature has been used to screen for suppressors of an ABA-deficient mutant (aba3-1) cold-leaf phenotype. Three novel mutants, called hot ABA-deficiency suppressor (has), have been identified with hot-leaf phenotypes in the absence of the aba3 mutation. The defective genes imparted no apparent modification to ABA production on water deficit, were inherited recessively and enhanced ABA responses indicating that the proteins encoded are negative regulators of ABA signalling. All three mutants showed ABA-hypersensitive stomata closure and inhibition of root elongation with little modification of growth and development in non-stressed conditions. The has2 mutant also exhibited increased germination inhibition by ABA, while ABA-inducible gene expression was not modified on dehydration, indicating the mutated gene affects early ABA-signalling responses that do not modify transcript levels. In contrast, weak ABA-hypersensitivity relative to mutant developmental phenotypes suggests that HAS3 regulates drought responses by both ABA-dependent and independent pathways. has1 mutant phenotypes were only apparent on stress or ABA treatments, and included reduced water loss on rapid dehydration. The HAS1 locus thus has the required characteristics for a targeted approach to improving resistance to water deficit. In contrast to has2, has1 exhibited only minor changes in susceptibility to Dickeya dadantii despite similar ABA-hypersensitivity, indicating that crosstalk between ABA responses to this pathogen and drought stress can occur through more than one point in the signalling pathway

ABSCISIC ACID-DEFICIENT4 Has an Essential Function in Both cis-Violaxanthin and cis-Neoxanthin Synthesis

International audienceAbscisic acid (ABA), a plant hormone synthesized from carotenoids, functions in seed germination and abiotic stress responses. ABA is derived from the cleavage of 9-cis-isomers of violaxanthin and neoxanthin, which are oxygenated carotenoids, also called xanthophylls. Although genes encoding enzymes responsible for most steps of the ABA biosynthesis pathway have been identified, enzymatic reactions leading to the production of these cis-isomers from trans-violaxanthin remain poorly understood. Two mutants that lack trans- and cis-neoxanthin, tomato (Solanum lycopersicum) neoxanthin-deficient1 (nxd1) and Arabidopsis (Arabidopsis thaliana) ABA-deficient4 (aba4), were identified previously, but only aba4 exhibited ABA-deficient phenotypes. No enzymatic activity was detected for ABA4 and NXD1 proteins, and their exact function remained unknown. To further investigate ABA4 and NXD1 function in Arabidopsis, we compared phenotypes of single and double mutants, and analyzed the effect of ABA4 overexpression on ABA and carotenoid accumulation in wild-type and mutant backgrounds. We provide convergent evidence that ABA4 is not only required for the formation of trans- and 9 '-cis-neoxanthin from trans-violaxanthin, but also controls 9-cis-violaxanthin accumulation. While nxd1 produces high amounts of 9-cis-violaxanthin and ABA, aba4 nxd1 exhibits reduced levels in both leaves and seeds. Furthermore, ABA4 constitutive expression in nxd1 increases both 9-cis-violaxanthin and ABA accumulation. Subcellular localization of NXD1 protein in transient expression assays suggests that production of the NXD1-derived factor required for neoxanthin synthesis takes place in the cytosol. Finally, we postulate that ABA4, with additional unknown cofactor(s), is required for, or contributes to, trans-to-cis violaxanthin isomerase activity, producing both cis-xanthophyll precursors of ABA

Deciphering the molecular mechanisms underpinning the transcriptional control of gene expression by L-AFL proteins in Arabidopsis seed.

International audienceIn Arabidopsis (Arabidopsis thaliana), transcriptional control of seed maturation involves three related regulators with a B3domain, namely LEAFY COTYLEDON2 (LEC2), ABSCISIC ACID INSENSITIVE3 (ABI3), and FUSCA3 (ABI3/FUS3/LEC2[AFLs]). Although genetic analyses have demonstrated partially overlapping functions of these regulators, the underlyingmolecular mechanisms remained elusive. The results presented here confirmed that the three proteins bind RY DNA elements(with a 59-CATG-39 core sequence) but with different specificities for flanking nucleotides. In planta as in the moss Physcomitrellapatens protoplasts, the presence of RY-like (RYL) elements is necessary but not sufficient for the regulation of the OLEOSIN1 (OLE1)promoter by the B3AFLs. G box-like domains, located in the vicinity of the RYL elements, also are required for proper activation ofthe promoter, suggesting that several proteins are involved. Consistentwith this idea, LEC2 andABI3 showed synergistic effects onthe activation of the OLE1 promoter. What is more, LEC1 (a homolog of the NF-YB subunit of the CCAAT-binding complex)further enhanced the activation of this target promoter in the presence of LEC2 and ABI3. Finally, recombinant LEC1 and LEC2proteins produced in Arabidopsis protoplasts could form a ternary complex with NF-YC2 in vitro, providing a molecular explanationfor their functional interactions. Taken together, these results allow us to propose a molecularmodel for the transcriptional regulationof seed genes by the L-AFL proteins, based on the formation of regulatory multiprotein complexes between NF-YBs, which carry aspecific aspartate-55 residue, and B3 transcription factors